CN102023181A - Enzyme electrode and preparation method thereof - Google Patents
Enzyme electrode and preparation method thereof Download PDFInfo
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- CN102023181A CN102023181A CN2009100938088A CN200910093808A CN102023181A CN 102023181 A CN102023181 A CN 102023181A CN 2009100938088 A CN2009100938088 A CN 2009100938088A CN 200910093808 A CN200910093808 A CN 200910093808A CN 102023181 A CN102023181 A CN 102023181A
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- 102000004190 Enzymes Human genes 0.000 title claims abstract description 84
- 108090000790 Enzymes Proteins 0.000 title claims abstract description 84
- 238000002360 preparation method Methods 0.000 title claims abstract description 25
- 239000004065 semiconductor Substances 0.000 claims abstract description 19
- 229940088598 enzyme Drugs 0.000 claims description 76
- 239000002070 nanowire Substances 0.000 claims description 53
- 239000000243 solution Substances 0.000 claims description 24
- 239000004054 semiconductor nanocrystal Substances 0.000 claims description 23
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 claims description 16
- 229910052710 silicon Inorganic materials 0.000 claims description 16
- 239000010703 silicon Substances 0.000 claims description 16
- 229910052799 carbon Inorganic materials 0.000 claims description 15
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- 239000000463 material Substances 0.000 claims description 11
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- 235000019420 glucose oxidase Nutrition 0.000 claims description 8
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- 108010029541 Laccase Proteins 0.000 claims description 6
- 102000003425 Tyrosinase Human genes 0.000 claims description 6
- 108060008724 Tyrosinase Proteins 0.000 claims description 6
- 229910052787 antimony Inorganic materials 0.000 claims description 6
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 6
- 229910052737 gold Inorganic materials 0.000 claims description 6
- 239000010931 gold Substances 0.000 claims description 6
- 239000002994 raw material Substances 0.000 claims description 6
- 239000010439 graphite Substances 0.000 claims description 5
- 229910002804 graphite Inorganic materials 0.000 claims description 5
- 229910052748 manganese Inorganic materials 0.000 claims description 5
- 229910004613 CdTe Inorganic materials 0.000 claims description 4
- 229920000557 Nafion® Polymers 0.000 claims description 4
- 102000004316 Oxidoreductases Human genes 0.000 claims description 4
- 108090000854 Oxidoreductases Proteins 0.000 claims description 4
- 102000003992 Peroxidases Human genes 0.000 claims description 4
- 229910052782 aluminium Inorganic materials 0.000 claims description 4
- 229910052980 cadmium sulfide Inorganic materials 0.000 claims description 4
- 239000001257 hydrogen Substances 0.000 claims description 4
- 229910052739 hydrogen Inorganic materials 0.000 claims description 4
- 125000004435 hydrogen atom Chemical class [H]* 0.000 claims description 4
- 229910052738 indium Inorganic materials 0.000 claims description 4
- AMGQUBHHOARCQH-UHFFFAOYSA-N indium;oxotin Chemical group [In].[Sn]=O AMGQUBHHOARCQH-UHFFFAOYSA-N 0.000 claims description 4
- 229910052757 nitrogen Inorganic materials 0.000 claims description 4
- 108040007629 peroxidase activity proteins Proteins 0.000 claims description 4
- 229910052718 tin Inorganic materials 0.000 claims description 4
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- 238000012360 testing method Methods 0.000 description 8
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 6
- 239000001301 oxygen Substances 0.000 description 6
- 229910052760 oxygen Inorganic materials 0.000 description 6
- 239000008363 phosphate buffer Substances 0.000 description 6
- 238000011160 research Methods 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 108010093096 Immobilized Enzymes Proteins 0.000 description 2
- 241001481789 Rupicapra Species 0.000 description 2
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 2
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- BASFCYQUMIYNBI-UHFFFAOYSA-N platinum Chemical compound [Pt] BASFCYQUMIYNBI-UHFFFAOYSA-N 0.000 description 2
- 238000005498 polishing Methods 0.000 description 2
- 239000002002 slurry Substances 0.000 description 2
- 238000004506 ultrasonic cleaning Methods 0.000 description 2
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- -1 1-butyl-3-methylimidazole-tetrafluoroborate Chemical compound 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
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- Enzymes And Modification Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses an enzyme electrode which belongs to the technical field of biosensors. The enzyme electrode consists of a base electrode and a responding layer fixed on the surface of the base electrode, wherein the responding layer consists of doped semiconductor nanometer wires on which enzyme is fixed. The invention also discloses a preparation method of the enzyme electrode. The preparation method comprises the step of adopting the doped semiconductor nanometer wires to fix enzyme molecules so as to achieve the direct electrochemical performance of the enzyme molecules. The enzyme electrode has better catalytic activity and wider linear range.
Description
Technical field
The invention belongs to field of biosensors, be specifically related to a kind of enzyme electrode and preparation method thereof.
Background technology
Biology sensor has important practical value as a kind of analytical equipment in environmental monitoring, clinical examination, food and fields such as medicine analysis and biochemical analysis.Wherein owing to the enzyme as biocatalyst has efficient, single-minded excellent specific property, making becomes the main flow of biology sensor research based on the electrochemica biological sensor of enzyme base.
Three phases has mainly been experienced in the development of electrochemica biological sensor: first stage is with the first generation biology sensor of oxygen as the electric transmission mediator, and the concentration of measured object is measured in the consumption by oxygen in the detection architecture or the increase of hydrogen peroxide.Because it is influenced by partial pressure of oxygen and H
2O
2The overpotential height, disturb many, be subjected to shortcoming such as oxygen solubility restriction, having occurred with artificial mediator is that the redox mediator is constructed enzyme electrode, has solved the dependence problem of sensor to oxygen.But discharge slow and potential shortcomings such as toxicity because artificial mediator exists, limited its possibility of surveying in health check-up.The third generation sensor of broad research then is to utilize enzyme itself and interelectrode direct electron to shift the conversion of finishing signal at present, need not to introduce mediator, and is irrelevant with oxygen or other electron accepters, improved the performance of sensor greatly.
Realize that more effective direct electron shifts between enzyme and the electrode, will construct a suitable membrane electrode interface, and in the structure at this membrane electrode interface, it is most important that the selection of material just seems.Have only that those biocompatibilities are relatively good to be only first-selection at favorable material aspect the direct electron transfer between enzyme and the electrode again.
In recent years along with the continuous progress of nanosecond science and technology, nano material is in aviation, the energy, and fields such as biology have obtained using widely.Because nano material has great specific surface area, its application aspect biology sensor has obtained a lot of achievements in research, noble metal nano particles for example, material with carbon element, semiconductor nano material etc.Yet in electrochemica biological sensor, still exist the problem of effective raising enzyme molecular activity center and interelectrode electron transfer efficiency.Wherein mostly the application in biology sensor is to concentrate on the performance of material biology sensor as the enzyme molecular vehicle time of research different-shape about semiconductor nano material.The character of carrier material own to the research of the influence of enzymatic activity then seldom.The research of adopting the doped semiconductor nanocrystal line to change enzymatic activity is not also reported.Doping is as a kind of important method that changes semiconductor material character, after mixing different elements, and the electric conductivity of semiconductor material, carrier concentration, significant change can take place in character such as optical characteristics.Therefore, after doping elements has improved the conductance of semiconductor nanowires, when the immobilized enzyme biomolecule, can obviously improve electron transfer rate, strengthen sensor performance.
Summary of the invention
The object of the present invention is to provide a kind of enzyme electrode, this electrode utilize the doped semiconductor nanocrystal line fixedly plurality of enzymes realize that the direct electron between effectively fixing and the enzyme and the electrode of enzyme molecule shifts, change the character of material by synthetic semiconductor nanowires is mixed, and then reach the effect that improves enzyme Journal of Molecular Catalysis activity.
Another object of the present invention is to provide the preparation method of above-mentioned enzyme electrode.
A kind of enzyme electrode, be made of basal electrode and the responding layer that anchors at the basal electrode surface, described responding layer is the doped semiconductor nanocrystal line that is fixed with enzyme, and described basal electrode is indium tin oxide (ITO), glass-carbon electrode, the pyrolytic graphite electrode, carbon paste electrode or gold electrode; Described enzyme is horseradish peroxidase, haemoglobin, myoglobins, cromoci, glucose oxidase, tyrosinase, laccase, reductase, hydrogen peroxidase, dehydrogenasa or oxidoreducing enzyme; The material of described semiconductor nanowires is ZnO, SnO
2, In
2O
3, Cu
2O, WO
3, Fe
3O
4, TiO
2, CdTe or CdS; Dopant in the described doped semiconductor nanocrystal line is Sb, In, Sn, Al, N, Ga, La, Mn or Co.
The mass ratio of the dopant in the described doped semiconductor nanocrystal line and the raw material of semiconductor nanowires is (5-20): 100.
To achieve these goals, the present invention adopts semiconductor nanowires to mix a method that is coated with zymoprotein, prepares enzyme electrode, and to have realized the Direct Electrochemistry behavior of zymoprotein, its concrete operations step is as follows:
1) prepare the doped semiconductor nanocrystal line with thermal evaporation method the silicon chip of gold-plated or ITO is put into the magnetic boat that raw material is housed, the mass ratio of the raw material of dopant and semiconductor nanowires is (5-20): 100, and at 700-950 ℃ of N with 50-800sccm
2Under the flow conditions, heat and to obtain the doped semiconductor nanocrystal line at silicon chip surface in 1～3 hour.
2) preparation of enzyme electrode
With diameter is that the basal electrode of 3mm is the Al of 1.0,0.3 and 0.05 μ m successively with diameter
2O
3Slurry is polished to minute surface on chamois leather, ultrasonic washing with clean water 2～3min is used then with clear water flush away surface contaminants in each polishing back, uses each ultrasonic cleaning 2～3min of ethanol and distilled water at last successively, N
2Dry up.The doped semiconductor nanocrystal line of step 1) preparation is placed solvent, make its even dispersion, described nano wire concentration is 1～10mg/mL; Enzyme is dissolved in the damping fluid, and described enzyme concentration is 1～10mg/mL; Above-mentioned doped semiconductor nanocrystal line solution and enzyme solutions are mixed, wherein, the volume ratio of doped semiconductor nanocrystal line solution and enzyme solutions is 1: (0.5-2), get mixed solution 6～10 μ L and drip the basal electrode surface that is polished to minute surface that is coated in above-mentioned cleaning, refrigerator dries for 4 ℃.
Described enzyme is horseradish peroxidase (HRP), haemoglobin, myoglobins, cromoci, glucose oxidase, tyrosinase, laccase, reductase, hydrogen peroxidase, dehydrogenasa or oxidoreducing enzyme.
Described basal electrode is indium tin oxide (ITO), glass-carbon electrode, pyrolytic graphite electrode, carbon paste electrode or gold electrode.
The material of described semiconductor nanowires is ZnO, SnO
2, In
2O
3, Cu
2O, WO
3, Fe
3O
4, TiO2, CdTe or CdS.
Dopant in the described doped semiconductor nanocrystal line is Sb, In, Sn, Al, N, Ga, La, Mn or Co.
Described doped semiconductor nanocrystal line is dispersed in the room-temperature ion liquid liquid solution of the acetum of ethanolic solution, 0.1～0.5wt% shitosan of absolute ethyl alcohol, 0.5～2wt%Nafion or BF-4 series.Wherein, the concentration of acetic acid is 0.01～0.1M in the described acetum.
Beneficial effect of the present invention: adopt doped semiconductor nanocrystal line immobilized enzyme molecule, make the enzyme molecule keep good active and have good catalytic property, the sensor of preparation has the very wide range of linearity, very high sensitivity and good stability, has still kept about 90% activity in test after month.
Description of drawings
Fig. 1 is nano wire/enzyme electrode synoptic diagram;
The 1-basal electrode; The 2-enzyme; 3-doped semiconductor nanocrystal line
Fig. 2 is the H with variable concentrations
2O
2The figure as a result that the enzyme electrode of embodiment 1 is tested as substrate.
The enzyme electrode of A: embodiment 1 preparation is to the H of variable concentrations
2O
2The current-responsive curve
B: the matched curve of electric current-concentration
Embodiment
Below in conjunction with Fig. 1 nano wire/enzyme electrode of the present invention is described.Enzyme electrode of the present invention comprises basal electrode 1 and anchors at the responding layer on basal electrode surface that responding layer is the doped semiconductor nanocrystal line 3 that is fixed with enzyme 2.
Below for the preparation method embodiment of enzyme electrode of the present invention.
The basal electrode that uses in following examples is in the disposal route of dripping before being coated with: is the Al of 1.0,0.3 and 0.05 μ m with diameter successively with diameter as the basal electrode of 3mm
2O
3Slurry is polished to minute surface on chamois leather, ultrasonic washing with clean water 2～3min is used then with clear water flush away surface contaminants in each polishing back, uses each ultrasonic cleaning 2～3min of ethanol and distilled water at last successively, N
2Dry up.
The SnO that preparation Sb mixes
2Nano wire: gold-plated silicon chip is put into the magnetic boat that glass putty and antimony powder are housed, and wherein, the mass ratio of glass putty and antimony powder is 100: 5, at 850 ℃ of N with 80sccm
2Flow conditions heating down can obtain the SnO that Sb mixes at silicon chip surface in 2 hours
2Nano wire;
To contain the above-mentioned Sb doping of 1mg/mL SnO
2The ethanolic solution of nano wire evenly mixes with the phosphate buffer equal-volume that contains 2mg/mL HRP, gets 6 μ L and drips and be coated in the glass-carbon electrode surface of handling through said method, and refrigerator dries for 4 ℃, thereby prepares nano wire/HRP laminated film enzyme electrode.
Adopt three-electrode system, above-mentioned enzyme electrode is tested, wherein platinum electrode is to electrode, and Ag/AgCl (saturated KCl) electrode is a contrast electrode, and above-mentioned enzyme electrode is as working electrode, and substrate is H
2O
2Solution.The results are shown in Figure 2, in the time less than 5s, electric current has reached 95% of steady-state current, and the response time is very short.Response current and concentration have good linear relationship in 10～450 μ M scopes, and sensitivity is 100mAM
-1Cm
-2The result shows that fixing enzyme molecule shows good electro catalytic activity, and the sensor of preparation has the very wide range of linearity, very high sensitivity and well stable, and test has still kept 90% activity after one month.And be fixed on unadulterated SnO
2The sensitivity of the HRP enzyme electrode of nanowire surface only is 45mA M
-1Cm
-2, much smaller than the sensitivity of the sensor of the nano wire preparation of mixing.
Embodiment 2
Sb doping SnO with 2mg
2Nano wire is dispersed in the Nafion ethanolic solution of 1mL 1wt%, with the phosphate buffer solution that contains 6mg/mL myoglobins (Mb) is evenly to mix at 1: 0.9 by volume, getting 7 μ L drips and is coated in the gold electrode surfaces of handling through said method, refrigerator dries for 4 ℃, thereby prepares nano wire/Mb laminated film enzyme electrode.
According to the H of the method among the embodiment 1 with variable concentrations
2O
2As substrate the enzyme electrode of embodiment 2 is tested, sensitivity has good linear relationship in 1～600 μ M scope, and stability is fine, and test still keeps 91% activity after four weeks.
Embodiment 3
With the Sb SnO that mixes
2Nano wire is distributed in the acetum (acetate concentration 0.05M in the solution) that contains the 0.3wt% shitosan, wherein, and Sb doping SnO in the solution
2The concentration of nano wire is 1mg/mL, this solution and the phosphate buffer solution that contains the 2mg/mL cromoci were evenly mixed in 1: 1.25 by volume, get 8 μ L and drip and be coated in the pyrolytic graphite electrode surface that cleaned, refrigerator dries for 4 ℃, thereby prepares nano wire/cromoci laminated film enzyme electrode.
According to the H of the method among the embodiment 1 with variable concentrations
2O
2As substrate the enzyme electrode of embodiment 3 is tested, had the very wide range of linearity (10～400 μ M), very high sensitivity (90mA M
-1Cm
-2) and well stable, test has still kept 85% activity after one month.
Embodiment 4
With the Sb SnO that mixes
2Nano wire is distributed in 1-butyl-3-methylimidazole-tetrafluoroborate ionic liquid at room temperature, its nano wire concentration is 5mg/mL, this solution and the phosphate buffer solution that contains the 8mg/mL haemoglobin were evenly mixed in 1: 0.8 by volume, getting 6 μ L drips and is coated in the carbon paste electrode surface of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/haemoglobin laminated film enzyme electrode.
According to the H of the method among the embodiment 1 with variable concentrations
2O
2As substrate the enzyme electrode of embodiment 4 is tested, had the very wide range of linearity (1～750 μ M) and very high sensitivity (120mA M
-1Cm
-2).
To contain 5mg/mLSb doping SnO
2The ethanolic solution of nano wire evenly mixes with the phosphate buffer solution equal-volume that contains the 8mg/mL glucose oxidase, gets 6 μ L and drips and be coated in the ITO electrode surface that cleaned, and refrigerator dries for 4 ℃, thereby prepares nano wire/glucose oxidase laminated film enzyme electrode.
As substrate the enzyme electrode of embodiment 5 is tested with the glucose of variable concentrations according to the method among the embodiment 1, (0.05～8mM) and well stable, test has still kept 89% activity after one month to have the very wide range of linearity.
Embodiment 6
The ZnO nano wire that preparation Al mixes: gold-plated silicon chip is put into the magnetic boat that Al powder and Zn powder are housed, wherein, the mass ratio of Zn powder and Al powder is 100: 15, under the stream of nitrogen gas condition of 900 ℃ and 400sccm, heated 2 hours, and can obtain the ZnO nano wire that Al mixes at silicon chip surface;
The ZnO nano wire that Al is mixed is distributed in the ethanol solution, the concentration of its nano wire is 1mg/mL, this solution is evenly mixed with volume ratio with the phosphate buffer that contains 2mg/mL Mb at 1: 1.25, getting 6 μ L drips and is coated in the glass-carbon electrode surface of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/Mb laminated film enzyme electrode.
According to the H of the method among the embodiment 1 with variable concentrations
2O
2As substrate the enzyme electrode of embodiment 6 is tested, had the very wide range of linearity (5～600 μ M) and very high sensitivity (90mAM
-1Cm
-2).
Embodiment 7
The In that preparation Sn mixes
2O
3Nano wire: the silicon chip that will be coated with ITO is put into the magnetic boat that In powder and Sn powder are housed, and wherein, the mass ratio of Sn powder and In powder is 9: 100, under the stream of nitrogen gas condition of 900 ℃ and 400sccm, heat 2 hours, can obtain the In of Sn doping at silicon chip surface
2O
3Nano wire;
To contain the In that 5mg/mLSn mixes
2O
3The ethanolic solution of the 2wt%Nafion of nano wire evenly mixed with the phosphate buffer that contains the 2mg/mL laccase in 1: 1.2 by volume, getting 6 μ L drips and is coated in the gold electrode surfaces of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/laccase laminated film enzyme electrode.
Embodiment 8
Prepare the ZnO nano wire that In mixes: gold-plated silicon chip is put into In is housed
2O
3In the magnetic boat of powder and Zn powder, wherein, In
2O
3With the mass ratio of Zn powder be 20: 100, heating can obtain the ZnO nano wire that In mixes at silicon chip surface in 1 hour under 850 ℃ and 300sccm flow conditions;
The ethanolic solution that will contain the ZnO nano wire of 8mg/mL In doping evenly mixes with the phosphate buffer equal-volume that contains the 5mg/mL tyrosinase, getting 6 μ L drips and is coated in the glass-carbon electrode surface of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/tyrosinase laminated film enzyme electrode.
As substrate the enzyme electrode of embodiment 8 is tested with the phenol of variable concentrations according to the method among the embodiment 1, had the very wide range of linearity (8～700 μ M), very high sensitivity (95mAM
-1Cm
-2) and well stable, test has still kept 85% activity after one month.
Embodiment 9
Prepare the ZnO nano wire that In mixes: gold-plated silicon chip is put into In is housed
2O
3In the magnetic boat of powder and Zn powder, wherein, In
2O
3With the mass ratio of Zn powder be 10: 100, heating can obtain the ZnO nano wire that In mixes at silicon chip surface in 1 hour under 850 ℃ and 400sccm flow conditions;
The ethanolic solution that will contain the ZnO nano wire of 5mg/mL In doping evenly mixes with the phosphate buffer equal-volume that contains 5mg/mLHRP, getting 6 μ L drips and is coated in the glass-carbon electrode surface of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/HRP laminated film enzyme electrode.
According to the H of the method among the embodiment 1 with variable concentrations
2O
2As substrate the enzyme electrode of embodiment 9 is tested, had the very wide range of linearity (8～700 μ M), very high sensitivity (95mA M
-1Cm
-2) and well stable, test has still kept 85% activity after one month.
Embodiment 10
The ZnO nano wire that preparation Mn mixes: gold-plated silicon chip is put into the magnetic boat that Mn powder and Zn powder are housed, wherein, the mass ratio of Mn and Zn powder is 10: 100, and heating can obtain the ZnO nano wire that Mn mixes at silicon chip surface in 2 hours under 900 ℃ and 300sccm flow conditions;
The ethanolic solution that will contain the ZnO nano wire that 10mg/mL In mixes and the phosphate buffer that contains the 5mg/mL glucose oxidase are to mix at 1: 2 by volume, getting 6 μ L drips and is coated in the glass-carbon electrode surface of cleaning, refrigerator dries for 4 ℃, thereby prepares nano wire/glucose oxidase laminated film enzyme electrode.
As substrate the enzyme electrode of embodiment 10 is tested with the glucose of variable concentrations according to the method among the embodiment 1, (0.05～10mM) and well stable, test has still kept 92% activity after one month to have the very wide range of linearity.
Claims (8)
1. an enzyme electrode is characterized in that, is made of basal electrode and the responding layer that anchors at the basal electrode surface, and described responding layer is the doped semiconductor nanocrystal line that is fixed with enzyme; Described basal electrode is indium tin oxide, glass-carbon electrode, pyrolytic graphite electrode, carbon paste electrode or gold electrode; Described enzyme is horseradish peroxidase, haemoglobin, myoglobins, cromoci, glucose oxidase, tyrosinase, laccase, reductase, hydrogen peroxidase, dehydrogenasa or oxidoreducing enzyme; The material of described semiconductor nanowires is ZnO, SnO
2, In
2O
3, Cu
2O, WO
3, Fe
3O
4, TiO
2, CdTe or CdS; Dopant in the described doped semiconductor nanocrystal line is Sb, In, Sn, Al, N, Ga, La, Mn or Co.
2. enzyme electrode according to claim 1 is characterized in that, the mass ratio of the dopant in the described doped semiconductor nanocrystal line and the raw material of semiconductor nanowires is (5-20): 100.
3. the preparation method of an enzyme electrode is characterized in that, comprises following operation steps:
1) prepare the doped semiconductor nanocrystal line with thermal evaporation: the silicon chip of gold-plated or ITO is put into the magnetic boat that raw material is housed, and the mass ratio of the raw material of dopant and semiconductor nanowires is (5-20): 100, and at 700-950 ℃ of N with 50-800sccm
2Under the flow conditions, heat and to obtain the doped semiconductor nanocrystal line at silicon chip surface in 1～3 hour.
2) preparation of enzyme electrode places solvent with the doped semiconductor nanocrystal line of step 1) preparation, makes its even dispersion, and the concentration of described doped semiconductor nanocrystal line is 1～10mg/mL; Enzyme is dissolved in the damping fluid, and the concentration of enzyme is 1～10mg/mL; Semiconductor nanowires solution and enzyme solutions that gained is mixed mix, wherein, the semiconductor nanowires solution that mixes and the volume ratio of enzyme solutions are 1: (0.5-2), get mixed solution 6～10 μ L and drip and be coated in the basal electrode surface of cleaning that is polished to minute surface, refrigerator dries for 4 ℃.
4. the preparation method of enzyme electrode according to claim 3, it is characterized in that described enzyme is horseradish peroxidase, haemoglobin, myoglobins, cromoci, glucose oxidase, tyrosinase, laccase, reductase, hydrogen peroxidase, dehydrogenasa or oxidoreducing enzyme.
5. the preparation method of enzyme electrode according to claim 3 is characterized in that, described basal electrode is indium tin oxide, glass-carbon electrode, pyrolytic graphite electrode, carbon paste electrode or gold electrode.
6. the preparation method of enzyme electrode according to claim 3 is characterized in that, the material of described semiconductor nanowires is ZnO, SnO
2, In
2O
3, Cu
2O, WO
3, Fe
3O
4, TiO
2, CdTe or CdS.
7. the preparation method of enzyme electrode according to claim 3 is characterized in that, the dopant in the described doped semiconductor nanocrystal line is Sb, In, Sn, Al, N, Ga, La, Mn or Co.
8. the preparation method of enzyme electrode according to claim 3, it is characterized in that, described solvent is the ethanolic solution of absolute ethyl alcohol, 0.5～2wt%Nafion, the acetum of 0.1～0.5wt% shitosan or the room-temperature ion liquid liquid solution of BF-4 series, wherein, the concentration of acetic acid is 0.01～0.1M in the described acetum.
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